西藏拉鲁湿地土壤酵母菌多样性及产胞外酶活性菌株分布特性研究
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摘要
采用平板稀释法从西藏拉鲁湿地土壤中分离酵母菌,开展城市湿地酵母菌多样性研究。结合经典分类法及26S rDNA D1/D2区域序列分析,对获得的酵母菌菌株进行系统分类,并进行产胞外酶活性菌株的皿内初筛。结果显示,分离得到的83株酵母菌分属于隐球菌属Cryptococcus、假丝酵母属Candida、原囊菌属Protomyces、红酵母属Rhodotorula及孢囊线黑粉酵母Cystofilobasidium等5个属16个种,其中胶红酵母Rhodotorula mucilaginosa相对丰度最高,约26%,其次为维多利亚隐球酵母Cryptococcus victoriae和山杨隐球酵母Cryptococcus aspenensis;多样性分析表明,LhWS6样点的酵母菌多样性最丰富,LhWS9样点则最低;产胞外酶活性菌株初筛结果表明,25℃下产脂肪酶活性菌株比例最高,为67%,而4℃下产淀粉酶活性菌株数最多,为49株。另外,低温条件下产淀粉酶、果胶酶、纤维素酶和几丁质酶的活性菌株比例均高于常温条件。结果表明拉鲁湿地土壤酵母菌总体种类较丰富,但每个样点的种类相对单一,说明湿地生境或人为活动一定程度影响到该生态环境酵母菌的种群分布,开发青藏高原耐低温、产胞外酶等活性菌株具有一定的应用前景。
In this paper,we isolated yeasts from soil of Lhalu Wetland using the dilution method. In addition,we used 26S rDNA D1/D2 domain sequence analysis and traditional methods to identify isolates and screened extracelluar enzyme activity strains in petri dish. 83 yeast isolates were identified as belonging to 16 species of 5 genera,including Cryptococcus,Candida,Protomyces,Rhodotorula and Cystofilobasidium. R. mucilaginosa was the most species in soil of the wetland,moreover,the relative abundance of Cryptococcus victoriae and Cryptococcus aspenensis was also high. Diversity indices showed that yeast isolates in the Lh WS6 site were most abundant,however,that of the Lh WS9 site were the lowest. The study showed that human activities had influences on spatial dynamics of yeast community in Lhalu Wetland. The test also manifested that the percentage of lipase activity strains was the highest at 25℃,however,the number of strains produced amylase most at 4℃,up to 49 isolates. Furthermore,the percentage of isolates producing amlyse,pectinase,cellulase and chitinase was higher in cold condition. Yeasts distributing in soil of Lhalu Wetland were abundant,but in single site,the kind was few,which indicated that the wetland habitat or human activity could effect population distribution,and it was important to exploit low temperature-tolerant or exoenzyme activity microorganism in Qinghai-Tibet plateau.
In this paper,we isolated yeasts from soil of Lhalu Wetland using the dilution method. In addition,we used 26S rDNA D1/D2 domain sequence analysis and traditional methods to identify isolates and screened extracelluar enzyme activity strains in petri dish. 83 yeast isolates were identified as belonging to 16 species of 5 genera,including Cryptococcus,Candida,Protomyces,Rhodotorula and Cystofilobasidium. R. mucilaginosa was the most species in soil of the wetland,moreover,the relative abundance of Cryptococcus victoriae and Cryptococcus aspenensis was also high. Diversity indices showed that yeast isolates in the Lh WS6 site were most abundant,however,that of the Lh WS9 site were the lowest. The study showed that human activities had influences on spatial dynamics of yeast community in Lhalu Wetland. The test also manifested that the percentage of lipase activity strains was the highest at 25℃,however,the number of strains produced amylase most at 4℃,up to 49 isolates. Furthermore,the percentage of isolates producing amlyse,pectinase,cellulase and chitinase was higher in cold condition. Yeasts distributing in soil of Lhalu Wetland were abundant,but in single site,the kind was few,which indicated that the wetland habitat or human activity could effect population distribution,and it was important to exploit low temperature-tolerant or exoenzyme activity microorganism in Qinghai-Tibet plateau.
引文
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[13] 普布,孙扩,张二豪,等. 拉鲁湿地夏季昆虫群落组成特征的初步研究[J]. 西藏大学学报(自然科学版),2013,28(1):7-11.
[14] 巴桑,次仁,普布,等. 拉鲁湿地国家级自然保护区鸟类资源及其保护对策[J],资源科学,2009,31(7):1238-1243.
[15] 范丽卿,土艳丽,李建川,等. 拉萨市拉鲁湿地鱼类现状与保护[J]. 资源科学,2011,33(9):1742-1749.
[16] 何建清,张格杰,岳海梅. 拉鲁湿地自然保护区放线菌组成分析及生物活性测定[J]. 微生物学杂志,2009(4):6-10.
[17] 德吉,吴小妮,索朗德庆,等. 拉鲁湿地土壤细菌多样性初探[J]. 西藏大学学报(自然科学版),2014,29(2):44-50,
[18] 徐美鑫,刘天明,相茂功,等. 青海东部土壤中酵母物种多样性研究[J]. 微生物学通报,2009,36(3):360-364.
[2] 孔维栋. 极地陆域微生物多样性研究进展[J]. 生物多样性,2013,21(4):456-467.
[3] Zhang Y L,Wang C L,Bai W Q,et al. Alpine wetlands in the Lhasa River Basin,China[J]. Journal of Geographical Sciences,2010,20(3):375-388.
[4] 刘银银,李峰,孙庆业,等. 湿地生态系统土壤微生物研究进展[J]. 应用与环境生物学报,2013,19(3):547-552.
[5] 黄薇,陈叙图. 拉鲁湿地的生态服务功能及其退化原由的分析[J]. 西部林业科学,2008,37(3):41-45.
[6] 拉多,John Birks,普布,等. 拉萨市拉鲁湿地植物群落与环境因素之间的关系及其分布格局[J]. 西藏大学学报(自然科学版),2009,24(1):15-20.
[7] 巴桑,普布,马正学,等. 西藏拉鲁湿地夏季和秋季浮游藻类群落特征[J]. 湿地科学,2012,10(4):404-416.
[8] 陈德来,马正学,普布,等. 拉萨拉鲁湿地夏季土壤动物的群落特征[J]. 动物学杂志,2011,46(5):1-7.
[9] 巴桑,黄香,普布,等. 拉鲁湿地夏秋季纤毛虫群落特征及其与水环境的关系[J]. 生态学杂志,2014,33(10):2740-2748.
[10] 陈德来,普布,巴桑,等. 西藏拉鲁湿地夏季土壤线虫群落特征[J]. 动物学杂志,2014,49(5):744-753.
[11] 巴桑,普布. 拉鲁湿地自然保护区有壳肉足虫物种多样性与空间分布特征[J]. 西藏大学学报(自然科学版),2014,29(1):1-7.
[12] Chen L Y,Zhao X L,Ma H G,et al. Morphology,morphogenesis and molecular phylogeny of a soil ciliate Pseudouroleptus caudatus caudatus Hemberger,1985(Ciliophora,Hypotricha),from Lhalu Wetland,Tibet[J]. European Journal of Protistology,2015,51:1–14.
[13] 普布,孙扩,张二豪,等. 拉鲁湿地夏季昆虫群落组成特征的初步研究[J]. 西藏大学学报(自然科学版),2013,28(1):7-11.
[14] 巴桑,次仁,普布,等. 拉鲁湿地国家级自然保护区鸟类资源及其保护对策[J],资源科学,2009,31(7):1238-1243.
[15] 范丽卿,土艳丽,李建川,等. 拉萨市拉鲁湿地鱼类现状与保护[J]. 资源科学,2011,33(9):1742-1749.
[16] 何建清,张格杰,岳海梅. 拉鲁湿地自然保护区放线菌组成分析及生物活性测定[J]. 微生物学杂志,2009(4):6-10.
[17] 德吉,吴小妮,索朗德庆,等. 拉鲁湿地土壤细菌多样性初探[J]. 西藏大学学报(自然科学版),2014,29(2):44-50,
[18] 徐美鑫,刘天明,相茂功,等. 青海东部土壤中酵母物种多样性研究[J]. 微生物学通报,2009,36(3):360-364.
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